So how does it work? Their protocol uses mouse Tet (Ten-eleven translocation)-1, (or mTet1) enzyme expressed in a baculovirus system. First 5hmC is protected with a glucose using Beta-GT. Next, in a “One pot” procedure, the mTet1 converts 5hmC from 5mC, which is then immediately glycoslated by Beta-GT with a modified glucose moiety(6-N3-glucose) . Then all the original 5mC is labeled with biotin via click chemistry. Time for affinity biotin/streptavidin purification, followed by sequencing.

The researchers used mouse embryotic stem cells, as well as human breast cancer cell lines, to compare methylome data with other methods. TamC-Seq was efficient at providing more methylation site coverage than MeDIP-Seq. TamC-Seq captured a wider range of CpGs, showing less density bias than MeDIP-Seq. The TamC-Seq data was concordant with Bisulfite-Seq data.

The same research group is also working on RNA epigenetics. Interesting! Check out their web site.